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1.  Frequency of sucrose exposure on the cariogenicity of a biofilm-caries model 
European Journal of Dentistry  2016;10(3):345-350.
Although sucrose is considered the most cariogenic carbohydrate in the human diet, the question of how many exposures are needed to induce damage on the hard dental tissues remains unclear. To approach this question, different frequencies of daily sucrose exposure were tested on a relevant biological caries model.
Materials and Methods:
Biofilms of the Streptococcus mutans were formed on enamel slabs and exposed to cariogenic challenges with 10% sucrose for 5 min at 0, 1, 3, 5, 8, or 10 times per day. After 5 days, biofilms were retrieved to analyze biomass, protein content, viable bacteria, and polysaccharide formation. Enamel demineralization was evaluated by percentage of microhardness loss (percentage surface hardness loss [%SHL]).
Biomass, protein content, polysaccharide production, acidogenicity of the biofilm, and %SHL proportionally increased with the number of daily exposures to sucrose (P < 0.05). One daily sucrose exposure was enough to induce 20% more demineralization than the negative unexposed control. Higher frequencies induced greater demineralization and more virulent biofilms, but eight and ten exposures were not different between them in most of the analyzed variables (P > 0.05).
Higher sucrose exposure seems to increase cariogenicity, in a frequency-dependent manner, by the modification of bacterial virulent properties.
PMCID: PMC4926586  PMID: 27403051
Caries; diet; nutrition; Streptococcus mutans; sucrose; sugar
2.  Fiber optic backscatter spectroscopic sensor to monitor enamel demineralization and remineralization in vitro 
In this study, a Fiber Optic Backscatter Spectroscopic Sensor (FOBSS) is used to monitor demineralization and remineralization induced changes in the enamel. A bifurcated fiber optic backscatter probe connected to a visible light source and a high resolution spectrophotometer was used to acquire the backscatter light spectrum from the tooth surface. The experiments were conducted in two parts. In Part 1, experiments were carried out using fiber optic backscatter spectroscopy on (1) sound enamel and dentine sections and (2) sound tooth specimens subjected to demineralization and remineralization. In Part 2, polarization microscopy was conducted to examine the depth of demineralization in tooth specimens. The enamel and dentine specimens from the Part-1 experiments showed distinct backscatter spectra. The spectrum obtained from the enamel-dentine combination and the spectrum generated from the average of the enamel and dentine spectral values were closely similar and showed characteristics of dentine. The experiments in Part 2 showed that demineralization and remineralization processes induced a linear decrease and linear increase in the backscatter light intensity respectively. A negative correlation between the decrease in the backscatter light intensity during demineralization and the depth of demineralization determined using the polarization microscopy was calculated to be p = -0.994. This in vitro experiment highlights the potential benefit of using FOBSS to detect demineralization and remineralization of enamel.
PMCID: PMC2813093  PMID: 20142887
Fiber optic; demineralization; dentine; remineralization
3.  Effect of Fluoride on Artificial Caries Lesion Progression and Repair in Human Enamel: Regulation of Mineral Deposition and Dissolution under In Vivo‐Like Conditions 
Archives of oral biology  2006;52(2):110-120.
This study was carried out to determine in vitro the effect of fluoride on 1) the demineralization of sound human enamel and 2) the progression of artificial caries‐like lesions, under relevant oral conditions.
Thin sections of sound human enamel were exposed to solutions undersaturated with respect to tooth enamel to a degree similar to that found in dental plaque fluid following sucrose exposure in vivo, containing fluoride concentrations (0 – 0.38 ppm) found in plaque fluid. Mineral changes were monitored for 98 days, using quantitative microradiography. The effect of fluoride (1.0 – 25.0 ppm) on the progression of artificial caries‐like lesions was similarly studied.
Fluoride concentrations of 0.19 ppm and greater were found to prevent the demineralization of sound enamel in vitro. However, significantly higher concentrations of fluoride (25.0 ppm) were required to prevent further demineralization of artificial caries‐like lesions. Demineralizing solutions with intermediate fluoride concentrations (2.1 – 10.1 ppm) induced simultaneously remineralization in the outer portion of the lesion and demineralization in the inner portion. Simultaneous remineralization and demineralization were also observed in hydroxyapatite pellets.
Our results show that the observed effect of fluoride on enamel demineralization is not solely a function of bulk solution properties, but also depends on the caries‐status of the enamel surface. A mechanistic model presented indicates that, in comparison to sound enamel surfaces, higher concentrations of fluoride are required to prevent the progression of artificial caries‐like lesions under in vivo‐like conditions since the diffusion of mineral ions that promote remineralization is rate‐limiting.
PMCID: PMC2040327  PMID: 17049334
dental enamel; remineralization; demineralization; mineral distribution; fluoride
4.  Clinical Implications of Power Toothbrushing on Fluoride Delivery: Effects on Biofilm Plaque Metabolism and Physiology 
Dental biofilms are implicated in the formation of caries and periodontal disease. A major constituent of the supragingival biofilm is Streptococcus mutans, which produces lactic acid from sucrose fermentation, enhancing enamel demineralization and eventual caries development. Caries prevention through F inhibits enamel demineralization and promotes remineralization. Fluoride also exerts effects on metabolic activities in the supragingival biofilm such as aerobic respiration, acid fermentation and dentrification. In experimental S. mutans biofilms, adding 1000 ppm F to an acidogenic biofilm resulting from 10% sucrose addition increased pH to pre-sucrose levels, suggesting inhibition of acid fermentation. F effects on metabolic activity and sucrose utilization in interproximal plaque biofilms were also recorded. Addition of 10% sucrose reduced pH from neutral to 4.2, but subsequent addition of 1000 ppm F increased pH by 1 unit, inhibiting acid fermentation. 10% Sucrose addition also stimulated denitrification, increasing production of nitrous oxide (N2O). Addition of 1000 ppm F suppressed denitrification, indicating an additional mechanism by which F exerts effects in the active interproximal biofilm. Finally, fluid dynamic activity by power tooth brushing enhanced F delivery and retention in an experimental S. mutans biofilm, suggesting a potential novel benefit for this intervention beyond mechanical plaque removal.
PMCID: PMC2855952  PMID: 20414341
5.  Does erosion progress differently on teeth already presenting clinical signs of erosive tooth wear than on sound teeth? An in vitro pilot trial 
BMC Oral Health  2016;17:14.
Erosive tooth wear (ETW) is clinically characterized by a loss of tooth surface, and different enamel depths may have different susceptibility to demineralization. Therefore, the aim of this in vitro pilot study was to assess if the progression of erosive demineralization is faster on teeth already presenting signs of ETW when compared to originally sound teeth.
We selected 23 central incisors: 14 were clinically sound (Sound) and 9 presented clinical signs of early erosive tooth wear (ETW-teeth). The teeth were embedded in resin, leaving an uncovered window of native enamel (6.69 ± 2.30 mm2) on the incisal half of the labial surface. We measured enamel surface reflection intensity (SRI) initially and after each consecutive erosive challenge (1 % citric acid, total of 4, 8, 12, 16, 20 and 24 min). Calcium released to the citric acid was measured with an atomic absorption spectrometer.
We observed higher initial SRI values in ETW-teeth than in Sound teeth (p = 0.007). During in vitro erosive demineralization, we observed that erosion on originally Sound teeth progressed significantly slower (p = 0.033) than on ETW-teeth: SRI decreased by 75 % (from 100 to 25 %) on Sound teeth, and by 89 % (from 100 to 11 %) on ETW-teeth. Calcium release increased during erosion, but presented no significant differences (p = 0.643) between originally Sound (0.031 μmol/mm2) and ETW-teeth (0.032 μmol/mm2). There was satisfactory correlation between calcium release and rSRI values (rs = −0.66).
The optical reflectometer distinguished originally sound teeth from those with signs of ETW, and the results suggest that acid demineralization progresses differently on teeth already presenting clinical signs of ETW than on sound teeth.
Electronic supplementary material
The online version of this article (doi:10.1186/s12903-016-0231-y) contains supplementary material, which is available to authorized users.
PMCID: PMC4948097  PMID: 27430320
Dental erosion; Assessment; Enamel; Optical reflectometer; Reflection analysis; Diagnostics; Calcium; Susceptibility
6.  Atomic force microscopy study of enamel remineralization 
Annali di Stomatologia  2014;5(3):98-102.
The aim of the present in vitro study was the evaluation of two products: a CPP-ACP paste (GC Tooth Mousse, GC Corp.) and a desensitizing toothpaste (Colgate Sensitive Pro Relief, Colgate-Palmolive) on preventing enamel erosion produced by a soft drink (Coca Cola) by using Atomic Force Microscopy (AFM).
Thirty enamel specimens were assigned to 6 groups of 5 specimens each. 1: intact enamel, 2: enamel + soft drink, 3: intact enamel + Colgate Sensitive Pro Relief, 4: enamel + soft drink + Colgate Sensitive Pro Relief, 5: intact enamel + GC Tooth Mousse, 6: enamel + soft drink + GC Tooth Mousse. The surface of each specimen was imaged by AFM. The root mean-square roughness (Rrms) was obtained from the AFM images and the differences in the averaged values among the groups were analyzed by ANOVA test.
Comparing groups 4 and 6 (soft drink + toothpastes) with group 2 (eroded enamel) a statistical difference (P<0.05) was registered, suggesting effectiveness in protecting enamel against erosion of the products investigated.
The use of new formulation toothpastes can prevent enamel demineralization.
PMCID: PMC4252861  PMID: 25506414
AFM; enamel; remineralization; SEM; soft drinks; surface roughness
7.  Quantification of Dental Erosions in Patients With GERD Using Optical Coherence Tomography Before and After Double-Blind, Randomized Treatment With Esomeprazole or Placebo 
Dental erosion, the chemical dissolution of enamel without bacterial involvement, is a rarely reported manifestation of gastroesophageal reflux disease (GERD), as well as of recurrent vomiting and dietary habits. It leads to loss of tooth substance, hypersensitivity, functional impairment, and even tooth fracture. To date, dental erosions have been assessed using only very basic visual methods, and no evidence-based guidelines or studies exist regarding the prevention or treatment of GERD-related dental erosions.
In this randomized, double-blind study, we used optical coherence tomography (OCT) to quantify dental tissue demineralization and enamel loss before and after 3 weeks of acid-suppressive treatment with esomeprazole 20 mg b.i.d. or placebo in 30 patients presenting to the Berne University Dental Clinic with advanced dental erosions and abnormal acid exposure by 24-h esophageal pH manometry (defined as >4% of the 24-h period with pH < 4). Enamel thickness, reflectivity, and absorbance as measures of demineralization were quantified by OCT before and after therapy at identical localizations on teeth with most severe visible erosions as well as several other predefined changes in teeth.
The mean±s.e.m. decrease of enamel thickness of all teeth before and after treatment at the site of maximum exposure was 7.2±0.16 μm with esomeprazole and 15.25±0.17 μm with placebo (P =0.013), representing a loss of 0.3% and 0.8% of the total enamel thickness, respectively. The change in optical reflectivity to a depth of 25 μm after treatment was −1.122 ±0.769 dB with esomeprazole and +2.059±0.534 dB with placebo (P = 0.012), with increased reflectivity signifying demineralization.
OCT non-invasively detected and quantified significantly diminished progression of dental tissue demineralization and enamel loss after only 3 weeks of treatment with esomeprazole 20 mg b.i.d. vs. placebo. This suggests that esomeprazole may be useful in counteracting progression of GERD-related dental erosions. Further validation of preventative treatment regimens using this sensitive detection method is required, including longer follow-up and correlation with quantitative reflux measures.
PMCID: PMC4167766  PMID: 19654570
8.  Proteins, Pathogens, and Failure at the Composite-Tooth Interface 
Journal of Dental Research  2014;93(12):1243-1249.
In the United States, composites accounted for nearly 70% of the 173.2 million composite and amalgam restorations placed in 2006 (Kingman et al., 2012), and it is likely that the use of composite will continue to increase as dentists phase out dental amalgam. This trend is not, however, without consequences. The failure rate of composite restorations is double that of amalgam (Ferracane, 2013). Composite restorations accumulate more biofilm, experience more secondary decay, and require more frequent replacement. In vivo biodegradation of the adhesive bond at the composite-tooth interface is a major contributor to the cascade of events leading to restoration failure. Binding by proteins, particularly gp340, from the salivary pellicle leads to biofilm attachment, which accelerates degradation of the interfacial bond and demineralization of the tooth by recruiting the pioneer bacterium Streptococcus mutans to the surface. Bacterial production of lactic acid lowers the pH of the oral microenvironment, erodes hydroxyapatite in enamel and dentin, and promotes hydrolysis of the adhesive. Secreted esterases further hydrolyze the adhesive polymer, exposing the soft underlying collagenous dentinal matrix and allowing further infiltration by the pathogenic biofilm. Manifold approaches are being pursued to increase the longevity of composite dental restorations based on the major contributing factors responsible for degradation. The key material and biological components and the interactions involved in the destructive processes, including recent advances in understanding the structural and molecular basis of biofilm recruitment, are described in this review. Innovative strategies to mitigate these pathogenic effects and slow deterioration are discussed.
PMCID: PMC4237635  PMID: 25190266
dentin bonding agents; methacrylate; gp340; Streptococcus mutans; esterases; biofilm
9.  Multispectral near-IR reflectance imaging of simulated early occlusal lesions: Variation of lesion contrast with lesion depth and severity 
Lasers in surgery and medicine  2013;46(3):203-215.
Background and Objectives
Early demineralization appears with high contrast at near-IR wavelengths due to a ten to twenty fold difference in the magnitude of light scattering between sound and demineralized enamel. Water absorption in the near-IR has a significant effect on the lesion contrast and the highest contrast has been measured in spectral regions with higher water absorption. The purpose of this study was to determine how the lesion contrast changes with lesion severity and depth for different spectral regions in the near-IR and compare that range of contrast with visible reflectance and fluorescence.
Materials and Methods
Forty-four human molars were used in this in vitro study. Teeth were painted with an acid-resistant varnish, leaving a 4×4 mm window on the occlusal surface of each tooth exposed for demineralization. Artificial lesions were produced in the unprotected windows after 12–48 hr exposure to a demineralizing solution at pH-4.5. Near-IR reflectance images were acquired over several near-IR spectral distributions, visible light reflectance, and fluorescence with 405-nm excitation and detection at wavelengths greater than 500-nm. Crossed polarizers were used for reflectance measurements to reduce interference from specular reflectance. Cross polarization optical coherence tomography (CP-OCT) was used to non-destructively assess the depth and severity of demineralization in each sample window. Matching two dimensional CP-OCT images of the lesion depth and integrated reflectivity were compared with the reflectance and fluorescence images to determine how accurately the variation in the lesion contrast represents the variation in the lesion severity.
Artificial lesions appear more uniform on tooth surfaces exposed to an acid challenge at visible wavelengths than they do in the near-IR. Measurements of the lesion depth and severity using CP-OCT show that the lesion severity varies markedly across the sample windows and that the lesion contrast in the visible does not accurately reflect the large variation in the lesion severity. Reflectance measurements at certain near-IR wavelengths more accurately reflect variation in the depth and severity of the lesions.
The results of the study suggest that near-IR reflectance measurements at longer wavelengths coincident with higher water absorption are better suited for imaging early caries lesions.
PMCID: PMC3961544  PMID: 24375543
demineralization; dental caries; enamel; near-IR imaging; polarization
10.  A novel antibacterial orthodontic cement containing a quaternary ammonium monomer dimethylaminododecyl methacrylate 
Journal of dentistry  2014;42(9):1193-1201.
Demineralized lesions in tooth enamel around orthodontic brackets are caused by acids from cariogenic biofilm. This study aimed to develop a novel antibacterial orthodontic cement by incorporating a quaternary ammonium monomer dimethylaminododecyl methacrylate (DMADDM) into a commercial orthodontic cement, and to investigate the effects on microcosm biofilm response and enamel bond strength. DMADDM, a recently-synthetized antibacterial monomer, was incorporated into orthodontic cement at 0%, 1.5%, 3% and 5% mass fractions. Bond strength of brackets to enamel was measured. A microcosm biofilm model was used to measure metabolic activity, lactic acid production, and colony-forming units (CFU) on orthodontic cements. Shear bond strength was not reduced at 3% DAMDDM (p > 0.1), but was slightly reduced at 5% DMADDM, compared to 0% DMADDM. Biofilm viability was substantially inhibited when in contact with orthodontic cement containing 3% DMADDM. Biofilm metabolic activity, lactic acid production, and CFU were much lower on orthodontic cement containing DMADDM than control cement (p < 0.05). Therefore, the novel antibacterial orthodontic cement containing 3% DMADDM inhibited oral biofilms without compromising the enamel bond strength, and is promising to reduce or eliminate demineralization in enamel around orthodontic brackets.
PMCID: PMC4559222  PMID: 25035230
Antibacterial; microcosm biofilm; quaternary ammonium methacrylate; bond strength; lactic acid; metabolic activity
11.  Nondestructive Assessment of Early Tooth Demineralization Using Cross-Polarization Optical Coherence Tomography 
New methods are needed for the nondestructive measurement of tooth demineralization and remineralization to monitor the progression of incipient caries lesions (tooth decay) for effective nonsurgical intervention and to evaluate the performance of anticaries treatments such as chemical treatments or laser irradiation. Studies have shown that optical coherence tomography (OCT) has great potential to fulfill this role since it can be used to measure the depth and severity of early lesions with an axial resolution exceeding 10 µm, it is easy to apply in vivo and it can be used to image the convoluted topography of tooth occlusal surfaces. In this paper, a review of the use of polarization-sensitive-OCT for the measurement of tooth demineralization is provided along with some recent results regarding improved methods for the detection of caries lesions in the earliest stages of development. Automated methods of analysis were used to measure the depth and severity of demineralized bovine enamel produced using simulated caries models that emulate demineralization in the mouth. Significant differences in the depth and integrated reflectivity from the lesions were detected after only a few hours of demineralization. These results demonstrate that cross-polarization-OCT is ideally suited for the nondestructive assessment of early demineralization.
PMCID: PMC3109909  PMID: 21660217
Dental caries; polarization-sensitive-optical coherence tomography (PS-OCT); tooth demineralization
12.  A new hand-held optical reflectometer to measure enamel erosion: correlation with surface hardness and calcium release 
Scientific Reports  2016;6:25259.
In the present study, the surface reflection intensity (SRI) was measured from enamel with different induced erosion degrees using a hand-held pen-size reflectometer (Hand-Held) and a Table-Top reflectometer. To validate the Hand-Held reflectometer, we correlated its optical signals with the change of surface microhardness (SMH), and amount of calcium released from the enamel samples during erosion. We used 124 tooth enamel specimens that were exposed to an erosive challenge of either 1, 2, 4, 6, 8, or 10 minutes. SRI and SMH were measured before and after the erosive challenge and we also measured the amount of calcium released to the citric acid. Relative SRI loss (rSRIloss) and relative SMH loss (rSMHloss) were calculated. rSRIloss from the Hand-Held and the Table-Top reflectometers were similar and significantly correlated to rSMHloss and calcium release. The regression analyses showed a significant association between rSRIloss from both reflectometers and rSMHloss and calcium, showing that both reflectometers can be used to measure erosive demineralization of enamel. The Hand-Held reflectometer is capable of assessing in vitro erosion, correlating to other commonly used methods. It is small, easy to handle and provides fast measurement, being a possible candidate to measure erosion in clinical studies.
PMCID: PMC4848505  PMID: 27121129
13.  To Analyse the Erosive Potential of Commercially Available Drinks on Dental Enamel and Various Tooth Coloured Restorative Materials – An In-vitro Study 
With the enormous change in life style pattern of a common man through the past few decades, there has been proportional variation in the amount and frequency of consumption of drinks. An increased consumption of these drinks will concurrently increase enamel surface roughness by demineralization, resulting in hypersensitivity and elevated caries risk.
The present study was designed to evaluate the erosive potential of commercially available drinks on tooth enamel and various tooth coloured restorative materials.
Materials and Methods
Extracted human teeth were taken and divided into four groups i.e. tooth enamel, glass ionomer cement, composite and compomer. Four commercially available drinks were chosen these were Coca -Cola, Nimbooz, Frooti and Yakult. The pH of each drink was measured. Each group was immersed in various experimental drinks for a period of 14 days. The erosive potential of each drink was measured by calculating the change in average surface roughness of these groups after the immersion protocol in various drinks. The data analysis was done by One Way Anova, Post-Hoc Bonferroni, and paired t –test.
Group II-GIC showed highest values for mean of change in average surface roughness and the values were statistically significant (p<0.001) with tooth enamel, composite and compomer (p=0.002). Coca-cola showed the highest erosive potential and Yakult showed the lowest, there was no statistical significant difference between the results shown by Yakult and Frooti.
Characteristics which may promote erosion of enamel and tooth coloured restorative materials were surface texture of the material and pH of the drinks.
PMCID: PMC4948519  PMID: 27437343
Dental erosion; Immersion period; Surfcom; Surface roughness
14.  Imaging early demineralization with PS-OCT 
New methods are needed for the nondestructive measurement of tooth demineralization and remineralization to monitor the progression of incipient caries lesions (tooth decay) for effective nonsurgical intervention and to evaluate the performance of anti-caries treatments such as chemical treatments or laser irradiation. Studies have shown that optical coherence tomography (OCT) has great potential to fulfill this role since it can be used to measure the depth and severity of early lesions with an axial resolution exceeding 10-μm, it is easy to apply in vivo and it can be used to image the convoluted topography of tooth occlusal surfaces. In this paper we attempt to determine the earliest stage at which we can detect significant differences in lesion severity. Automated methods of analysis were used to measure the depth and severity of demineralized bovine enamel produced using a simulated caries model that emulates demineralization in the mouth. Significant differences in the depth and integrated reflectivity from the lesions were detected after only a few hours of demineralization. These results demonstrate that cross polarization OCT is ideally suited for the nondestructive assessment of early demineralization.
PMCID: PMC3293401  PMID: 22399835
polarization; optical coherence tomography; tooth demineralization; dental caries
15.  Methods for Monitoring Erosion Using Optical Coherence Tomography 
Since optical coherence tomography is well suited for measuring small dimensional changes on tooth surfaces it has great potential for monitoring tooth erosion. The purpose of this study was to explore different approaches for monitoring the erosion of enamel. Application of an acid resistant varnish to protect the tooth surface from erosion has proven effective for providing a reference surface for in vitro studies but has limited potential for in vivo studies. Two approaches which can potentially be used in vivo were investigated. The first approach is to measure the remaining enamel thickness, namely the distance from the tooth surface to the dentinal-enamel junction (DEJ). The second more novel approach is to irradiate the surface with a carbon dioxide laser to create a reference layer which resists erosion. Measuring the remaining enamel thickness proved challenging since the surface roughening and subsurface demineralization that commonly occurs during the erosion process can prevent resolution of the underlying DEJ. The areas irradiated by the laser manifested lower rates of erosion compared to the non-irradiated areas and this method appears promising but it is highly dependent on the severity of the acid challenge.
PMCID: PMC3898817  PMID: 24465117
erosion; optical coherence tomography; caries prevention; carbon dioxide laser
16.  Shedding New Light on Early Caries Detection 
Dental caries continues to be a common chronic disease among various population groups. Patient care can be improved with detection at the earliest stage. However, current techniques do not have sufficient sensitivity and specificity. We discuss 2 new methods — optical coherence tomography (OCT) and polarized Raman spectroscopy (PRS) — that are potentially useful for early caries detection and monitoring.
OCT produces morphologic depth images of near-surface tissue structures with a resolution that is an order of magnitude greater than ultrasound imaging. Based on measurement of back-scattered near infrared light, OCT shows that sound enamel causes high-intensity back-scattering at the tooth surface that decreases rapidly with depth. In contrast, incipient lesions cause higher light back-scattering at the tooth surface and subsurface scattering indicative of porosity caused by demineralization. The scatter region within the enamel correlates well with the classical triangular shape of subsurface lesions observed in histologic sections. OCT imaging not only allows identification of incipient lesions, but also provides information on surface integrity and lesion depth.
PRS furnishes biochemical information about the tooth's composition, mineral content and crystallinity. The depolarization ratio derived from the dominant phosphate peak of hydroxyapatite in sound teeth is consistently lower than that from incipient caries. This difference is attributed to the change in enamel crystallite morphology or orientation that occurs with acid demineralization. Thus, PRS can be used to confirm suspect lesions determined by OCT and rule out false-positive signals from non-carious anomalies.
The combination of OCT and PRS provides a new detection method with high sensitivity and specificity that will improve caries management and patient care. Future studies are aimed at developing intraoral probes to validate the findings in vivo.
PMCID: PMC2700544  PMID: 19126361
17.  A Method for Monitoring Enamel Erosion Using Laser Irradiated Surfaces and Optical Coherence Tomography 
Lasers in surgery and medicine  2014;46(9):672-678.
Since optical coherence tomography (OCT) is well suited for measuring small dimensional changes on tooth surfaces, OCT has great potential for monitoring tooth erosion. Previous studies have shown that enamel areas ablated by a carbon dioxide laser manifested lower rates of erosion compared to the non-ablated areas. The purpose of this study was to develop a model to monitor erosion in vitro that could potentially be used in vivo.
Thirteen bovine enamel blocks were used in this in vitro study. Each 10 mm × 2 mm block was partitioned into five regions, the central region was unprotected, the adjacent windows were irradiated by a CO2 laser operating at 9.3 μm with a fluence of 2.4J/cm2, and the outermost windows were coated with acid resistant varnish. The samples were exposed to a pH cycling regimen that caused both erosion and subsurface demineralization for 2, 4 and 6 days. The surfaces were scanned using a time-domain polarization sensitive optical coherence tomography (PS-OCT) system and the degree of surface loss (erosion) and the integrated reflectivity with lesion depth was calculated for each window.
There was a large and significant reduction in the depth of surface loss (erosion) and the severity of demineralization in the areas irradiated by the laser.
Irradiation of the enamel surface with a pulsed carbon dioxide laser at sub-ablative intensities results in significant inhibition of erosion and demineralization under the acid challenge employed in this study. In addition, these results suggest that it may be feasible to modify regions of the enamel surface using the laser to serve as reference marks to monitor the rate of erosion in vivo.
PMCID: PMC4675665  PMID: 25147133
tooth erosion; tooth demineralization; carbon dioxide laser; optical coherence tomography
18.  Validation of a Cariogenic Biofilm Model to Evaluate the Effect of Fluoride on Enamel and Root Dentine Demineralization 
PLoS ONE  2016;11(1):e0146478.
Due to gingival recession both enamel and root dentine are at risk of developing caries. Both tissues are exposed to a similar environment, however there is not a validated model to evaluate the effect of fluoride on these dental substrates simultaneously. Hence, this study aimed to validate a caries model to evaluate the effect of fluoride to prevent demineralization on enamel and root-dentine. Streptococcus mutans UA159 biofilms were formed on saliva-coated bovine enamel and root dentine slabs (n = 12 per group) mounted in the same well of culture plates. The biofilms were exposed 8×/day to 10% sucrose and treated 2×/day with fluoridated solutions containing 0, 150, 450, or 1,350 ppm F; thus, simulating the use of low to high fluoride concentration toothpastes. The pH values of the culture medium was monitored 2×/day as a biofilm acidogenicity indicator. After 96 h, biofilms were collected for fluoride concentration analysis. The percentage of surface hardness loss (%SHL) was calculated for slabs. The fluoride uptake by the enamel and dentine was also determined. The model showed a dose-response because the biofilm and fluoride uptake increased and %SHL decreased at increasing fluoride concentrations (p < 0.05). Fluoride in the biofilm formed on dentine and fluoride uptake by dentine were higher than those for enamel. With the same fluoride concentration treatment, the percentage of reduction of demineralization was lower for dentine than for enamel. In conclusion, the model was validated in terms of a dose-response effect of fluoride on enamel and root dentine. Furthermore, the findings support the clinical data, suggesting that higher fluoride concentrations are necessary to control caries of root dentine than of enamel.
PMCID: PMC4712139  PMID: 26731743
19.  Remineralization of Demineralized Enamel via Calcium Phosphate Nanocomposite 
Journal of Dental Research  2012;91(10):979-984.
Secondary caries remains the main problem limiting the longevity of composite restorations. The objective of this study was to investigate the remineralization of demineralized human enamel in vitro via a nanocomposite containing nanoparticles of amorphous calcium phosphate (NACP). NACP were synthesized by a spray-drying technique and incorporated into a dental resin. First, caries-like subsurface enamel lesions were created via an acidic solution. Then, NACP nanocomposite or a commercial fluoride-releasing control composite was placed on the demineralized enamel, along with control enamel without a composite. These specimens were then treated with a cyclic demineralization/remineralization regimen for 30 days. Quantitative microradiography showed typical enamel subsurface demineralization before cyclic demineralization/remineralization treatment, and significant remineralization in enamel under the NACP nanocomposite after the demineralization/remineralization treatment. The NACP nanocomposite had the highest enamel remineralization (mean ± SD; n = 6) of 21.8 ± 3.7%, significantly higher than the 5.7 ± 6.9% for fluoride-releasing composite (p < 0.05). The enamel group without composite had further demineralization of −26.1 ± 16.2%. In conclusion, a novel NACP nanocomposite was effective in remineralizing enamel lesions in vitro. Its enamel remineralization was 4-fold that of a fluoride-releasing composite control. Combined with the good mechanical and acid-neutralization properties reported earlier, the new NACP nanocomposite is promising for remineralization of demineralized tooth structures.
PMCID: PMC3446834  PMID: 22933607
dental nanocomposite; calcium phosphate nanoparticles; human enamel; lesion remineralization; contact microradiography; caries inhibition
20.  Effect of an Orphan Response Regulator on Streptococcus mutans Sucrose-Dependent Adherence and Cariogenesis  
Infection and Immunity  2003;71(8):4351-4360.
Streptococcus mutans is the principal acidogenic component of dental plaque that demineralizes tooth enamel, leading to dental decay. Cell-associated glucosyltransferases catalyze the sucrose-dependent synthesis of sticky glucan polymers that, together with glucan binding proteins, promote S. mutans adherence to teeth and cell aggregation. We generated an S. mutans Tn916 transposon mutant, GMS315, which is defective in sucrose-dependent adherence and significantly less cariogenic than the UA130 wild-type progenitor in germfree rats. The results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western blotting, and N-terminal sequence analysis confirmed the absence of a 155-kDa glucosyltransferase S (Gtf-S) from GMS315 protein profiles. Mapping of the unique transposon insertion in GMS315 revealed disruption of a putative regulatory region located upstream of gcrR, a gene previously described by Sato et al. that shares significant amino acid identity with other bacterial response regulators (Y. Sato, Y. Yamamoto, and H. Kizaki, FEMS Microbiol. Lett. 186: 187-191, 2000). The gcrR regulator, which we call “tarC,” does not align with any of the 13 proposed two-component signal transduction systems derived from in silico analysis of the S. mutans genome, but rather represents one of several orphan response regulators in the genome. The results of Northern hybridization and/or real-time reverse transcription-PCR experiments reveal increased expression of both Gtf-S and glucan binding protein C (GbpC) in a tarC knockout mutant (GMS900), thereby supporting the notion that TarC acts as a negative transcriptional regulator. In addition, we noted that GMS900 has altered biofilm architecture relative to the wild type and is hypocariogenic in germfree rats. Taken collectively, these findings support a role for signal transduction in S. mutans sucrose-dependent adherence and aggregation and implicate TarC as a potential target for controlling S. mutans-induced cariogenesis.
PMCID: PMC166011  PMID: 12874312
21.  High Contrast Near-infrared Polarized Reflectance Images of Demineralization on Tooth Buccal and Occlusal Surfaces at λ=1310-nm 
Lasers in surgery and medicine  2009;41(3):208-213.
Background and Objectives:
Sound enamel manifests peak transparency in the near-IR (NIR) at 1310-nm, therefore the near-IR is ideally suited for high contrast imaging of dental caries. The purpose of this study was to acquire images of early demineralized enamel on the buccal and occlusal surfaces of extracted human teeth using NIR reflectance imaging and compare the contrast of those images with the contrast of images taken using other methods.
Materials and Methods:
Fifteen human molars were used in this in vitro study. Teeth were painted with a clear acid-resistant varnish, leaving two 2×2 mm windows on the buccal and occlusal surfaces of each tooth for demineralization. Artificial lesions were produced in the exposed windows after a 2-day exposure to a demineralizing solution at pH-4.5. Lesions were imaged using NIR transillumination, NIR and visible light reflectance, and fluorescence imaging methods. Crossed polarizers were used where appropriate to improve contrast. Polarization sensitive optical coherence tomography (PS-OCT) was also used to non-destructively assess the depth and severity of demineralization in each sample window.
NIR reflectance imaging had the highest image contrast for both the buccal and occlusal groups and it was significantly higher contrast than visible light reflectance (P<0.05).
The results of the study suggest that NIR reflectance imaging is a promising new method for acquiring high contrast images of early demineralization on tooth surfaces.
PMCID: PMC2689647  PMID: 19291753
demineralization; dental caries; enamel; near-IR Imaging; polarization
22.  Polarization Sensitive Optical Coherence Tomographic Imaging of Artificial Demineralization on Exposed Surfaces of Tooth Roots 
Background and Objectives
The purpose of this study was to assess the potential of polarization sensitive optical coherence tomography (PS-OCT) to non-destructively measure the depth and severity of artificial demineralization on exposed root surfaces and measure the degree of inhibition by topical fluoride. Although PS-OCT imaging studies have demonstrated the utility of PS-OCT for imaging carious lesions on enamel and dentin surfaces the influence of the cementum layer that is present on intact root surfaces has not been investigated.
Materials and Methods
In this study, extracted human tooth roots were partitioned into three sections with one partition treated with topical fluoride, one partition protected from demineralization with acid resistant varnish, and one partition exposed to a demineralization solution, producing artificial lesions approximately 200-µm deep in root dentin. The lesion depth, remaining cementum thickness and the integrated reflectivity for lesion areas was measured with PS-OCT. These measurements were also compared with more established methods of measuring demineralization, namely transverse microradiography (TMR) and polarized light microscopy (PLM)
PS-OCT was able to measure a significant increase in the reflectivity between lesion areas and sound root surfaces. In contrast to dentin, the cementum layer manifests minimal reflectivity in the PS-OCT images allowing nondestructive measurement of the remaining cementum thickness. The reflectivity of the cementum layer did not increase significantly after substantial demineralization, however it did manifest considerable shrinkage in a fashion similar to dentin and that shrinkage could be measured with OCT.
This study demonstrates that PS-OCT can be used to measure demineralization non-destructively on root surfaces and assess inhibition of demineralization by anticaries agents.
PMCID: PMC2701248  PMID: 19167052
optical coherence tomography; polarization; root dentin; cementum; artificial lesions; caries inhibition; microradiography; topical fluoride
23.  Enamel Carious Lesion Development in Response to Sucrose and Fluoride Concentrations and to Time of Biofilm Formation: An Artificial-Mouth Study 
Journal of oral diseases  2014;2014:348032.
The aim of this study was to evaluate both sucrose and fluoride concentrations and time of biofilm formation on enamel carious lesions induced by an in vitro artificial-mouth caries model. For Study 1, biofilms formed by streptococci and lactobacilli were grown on the surface of human enamel slabs and exposed to artificial saliva containing 0.50 or 0.75 ppmF (22.5 h/d) and broth containing 3 or 5% sucrose (30 min; 3x/d) over 5 d. In Study 2, biofilms were grown in the presence of 0.75 ppmF and 3% sucrose over 3 and 9 days. Counts of viable cells on biofilms, lesion depth (LD), and the integrated mineral loss (IML) on enamel specimens were assessed at the end of the tested conditions. Counts of total viable cells and L. casei were affected by sucrose and fluoride concentrations as well as by time of biofilm formation. Enamel carious lesions were shallower and IML was lower in the presence of 0.75 ppmF than in the presence of 0.50 ppmF (P < 0.005). No significant effect of sucrose concentrations was found with respect to LD and IML (P > 0.25). Additionally, deeper lesions and higher IML were found after 9 d of biofilm formation (P < 0.005). Distinct sucrose concentrations did not affect enamel carious lesion development. The severity of enamel demineralization was reduced by the presence of the higher fluoride concentration. Additionally, an increase in the time of biofilm formation produced greater demineralization. Our results also suggest that the present model is suitable for studying aspects related to caries lesion development.
PMCID: PMC4319183  PMID: 25664342
24.  Clinical Efficacy of a Specifically Targeted Antimicrobial Peptide Mouth Rinse: Targeted Elimination of Streptococcus mutans and Prevention of Demineralization 
Caries Research  2011;45(5):415-428.
Streptococcus mutans, the major etiological agent of dental caries, has a measurable impact on domestic and global health care costs. Though persistent in the oral cavity despite conventional oral hygiene, S. mutans can be excluded from intact oral biofilms through competitive exclusion by other microorganisms. This suggests that therapies capable of selectively eliminating S. mutans while limiting the damage to the normal oral flora might be effective long-term interventions to fight cariogenesis. To meet this challenge, we designed C16G2, a novel synthetic specifically targeted antimicrobial peptide with specificity for S. mutans. C16G2 consists of a S. mutans-selective ‘targeting region’ comprised of a fragment from S. mutans competence stimulation peptide (CSP) conjoined to a ‘killing region’ consisting of a broad-spectrum antimicrobial peptide (G2). In vitro studies have indicated that C16G2 has robust efficacy and selectivity for S. mutans, and not other oral bacteria, and affects targeted bacteria within seconds of contact.
In the present study, we evaluated C16G2 for clinical utility in vitro, followed by a pilot efficacy study to examine the impact of a 0.04% (w/v) C16G2 rinse in an intra-oral remineralization/demineralization model.
Results and Conclusions
C16G2 rinse usage was associated with reductions in plaque and salivary S. mutans, lactic acid production, and enamel demineralization. The impact on total plaque bacteria was minimal. These results suggest that C16G2 is effective against S. mutans in vivo and should be evaluated further in the clinic.
PMCID: PMC3169368  PMID: 21860239
Antimicrobial; Antimicrobial peptide; Caries; Demineralization; Dental plaque; Lactic acid; Mouth rinse; Oral therapeutic; Selective antibiotic; Selective therapeutic; Specifically targeted antimicrobial peptide; Streptococcus mutans; Targeted antimicrobial
25.  Selective Removal of Demineralization Using Near Infrared Cross Polarization Reflectance and a Carbon Dioxide Laser 
Lasers can ablate/remove tissue in a non-contact mode of operation and a pulsed laser beam does not interfere with the ability to image the tooth surface, therefore lasers are ideally suited for integration with imaging devices for image-guided ablation. Laser energy can be rapidly and efficiently delivered to tooth surfaces using a digitally controlled laser beam scanning system for precise and selective laser ablation with minimal loss of healthy tissues. Under the appropriate irradiation conditions such laser energy can induce beneficial chemical and morphological changes in the walls of the drilled cavity that can increase resistance to further dental decay and produce surfaces with enhanced adhesive properties to restorative materials. Previous studies have shown that images acquired using near-IR transillumination, optical coherence tomography and fluorescence can be used to guide the laser for selective removal of demineralized enamel. Recent studies have shown that NIR reflectance measurements at 1470-nm can be used to obtain images of enamel demineralization with very high contrast. The purpose of this study was to demonstrate that image guided ablation of occlusal lesions can be successfully carried out using a NIR reflectance imaging system coupled with a carbon dioxide laser operating at 9.3-μm with high pulse repetition rates.
PMCID: PMC3865262  PMID: 24357906
enamel; CO2 laser; dental caries; demineralization; near-IR imaging; reflectance

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